title: [Experimental study on the isolated culture of osteocytes and identification of osteoblasts in rats].
abstract: OBJECTIVE
  To establish a more stable method to isolate osteocytes in vitro, and then to find the differences with osteoblast biological characteristics.
  
  
  METHODS
  Osteocytes and osteoblasts were isolated from the bone tissue of 3-day-old rats using sequential collagenase digestion. The cells were identified through cell morphology after 24 hours later. Alkaline phosphatase (ALP) kit was used to stain the first generation cells by Kaplow-way, the bone gla protein (BGP) of the cells were stained by immunocytochemitry. Measured ALP and computed its activity.
  
  
  RESULTS
  Osteocytes and osteoblasts showed obviously differences in cell morphology. Osteocytes were star-shaped or dendrite-shaped within more dendrites, while osteoblasts were spindle-shaped with short dendrites. Osteocytes were negative for ALP, but osteoblasts were positive; Osteocytes were more positive for BGP, and osteoblasts were less positive. The secretion of ALP in osteocytes was lower than that of osteoblasts.
  
  
  CONCLUSIONS
  Osteocytes can be isolated and cultured in vitro. These characteristics of osteocytes are apparently difference with those of osteoblasts.
authors.affiliation: The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China.
authors.forename: Xiao-Kang
authors.initials: XK
authors.lastname: Tang
authors.forename: Wan
authors.initials: W
authors.lastname: Cheng
authors.forename: Bing
authors.initials: B
authors.lastname: Xu
authors.forename: Hang
authors.initials: H
authors.lastname: Ying
authors.forename: Pei-Jian
authors.initials: PJ
authors.lastname: Tong
authors.forename: Lu-wei
authors.initials: LW
authors.lastname: Xiao
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D010006:Osteoblasts / Q000737:chemistry / Q000166:cytology*; D015675:Osteocalcin / Q000032:analysis; D010011:Osteocytes / Q000737:chemistry / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D015675:Osteocalcin; D000469:Alkaline Phosphatase